C. elegans investigators universally agree that the post-genome future of C. elegans research must rest on a genetics platform. With this proposal, we lay the foundation for that platform. Our goals are as follows: 1) We have received approximately 700 requests to knockout C. elegans of which about 149 have been filled, leaving a backlog of about 550 genes. We propose to knock out those genes that remain on this request list. 2) We will develop ways to increase productivity and reduce the post per knockout through incremental improvements to our present gene knockout protocol. In collaboration with members of the C. Elegans Gene Knockout Consortium, we will then knock out C, elegans genes with significant homology to human genes. This highly significant class includes about 7,200 genes. At the Oklahoma Medical Research Foundation, we will do between 3,500 and 4,000 of the 7,200 potential targets. We will achieve these goals using methods developed in our lab that allow for the recovery of deletion mutants at specified target genes. However, using these methods, we are able to detect only a small fraction of the many possible types of chemically induced mutations in C. elegans. Therefore, we also propose research directed at expanding the types of mutations that we can detect, including single nucleotide mutations. Our objectives are to develop relatively low cost methods that will be useful in a high-throughput genomics project so that we then can knock out or otherwise modify the function of every c. elegans gene. This work supports and promotes the powerful genetic methods available for the study of C. elegans and it likely will lead to research that, realistically, would not take place if the expense and effort to make such gene knockouts were to be borne exclusively by individual investigators. Further, our research should not only be relevant to high throughput reverse-genetics, but will also enable more penetrating genetic studies in individual laboratories.. The ready availability of our proposed resource will accelerate the investigation of protein functions as it relates to the development and physiology of C. elegans and to the homologous processes found in vertebrates.